The long-term objective of Mynosys Cellular Devices is to develop new semi-robotic microdevices that enable scientists to directly physically probe or manipulate individual neurons or subcellular processes for a variety of experimental and therapeutic purposes. Envisioned as sub- millimeter-sized devices that have nano and microscale functional features, these novel devices allow scientists to operate at the same length-scale as neurons and their processes, facilitating new research questions and paradigms that thus far have eluded investigators due to a lack of appropriate miniaturized tools. The Specific Aims of the current Phase I proposal are to develop two easy-to-use silicon-based micro/nanodevices, that when placed in the hands of researchers, will serve as fundamental microtools for a broad range of neuroscience research on axon and dendritic function, injury, and regeneration. These two microscale research tools are an optically clear nanoknife comprising of a nanoscale sharp edge that allows accurate harvesting or experimental injury of elemental units of neural function such as axons and dendrites. In addition, we will produce an axon nanocompressor that can deliver highly calibrated compressive forces onto single axons to study how common forms of crushing nerve injury affects neurobiological function at the fundamental axonal level. As axons and dendrites are the essential units responsible for cellular communication in the nervous system, there is a substantial researcher base investigating axonal and dendritic biology in health and disease, who will benefit significantly from new abilities to harvest, probe, or manipulate individual axonal and dendritic segments for study. A clinically relevant research area that will benefit from these novel microscale tools is neural trauma, where the inability of axons in the adult spinal cord and brain to regenerate after injury is a very significant health and socioeconomic problem as well as a major challenge for neuroscience. The ability to deliver highly precise, repeatable, and calibrated injury forces onto single axons will aid in advancing this area of investigation. The research plan incorporates engineering design, microfabrication, with rigorous device mechanical and biological testing. The basic instrument design leverages an earlier company-developed prototype nanoknife, with the addition of key features to enhance user convenience and device robustness. Silicon wafer batch fabrication is also exploited to deliver both microtools at a relatively low cost to minimize barriers for adoption. The development of this micro-instrumentation will lay the foundation for future device enhancements such as the integration of on-board actuation to automate execution of the cutting or compression strokes. In addition, the experience and knowledge gained from the proposed project will be useful in the future development of additional neuro-microdevices to help advance research. PUBLIC HEALTH RELEVANCE: The proposed micro-instrumentation provides neuroscientists with the ability to directly manipulate and interact with nerve cells and their processes, allowing them to assay cell function at an unprecedented small scale. As the answers to major outstanding questions of diseases and illnesses of the nervous system will come from a deeper understanding of the inner workings of the neuron and its components, this novel micro-instrumentation will help facilitate scientific investigation, deepen our understanding of disease mechanisms, and accelerate the search for potential therapies.